Objective:
1. Develop and validate molecular identification and genotyping resources for insect biocontrol fungi Beauveria spp. and Metarhizium.
2. Distinguish fungal pathogens from species with potential for the biological protection of plants through molecular systematics investigations of complex coelomycete genera, including Colletotrichum and Pestalotiopsis.

Approach:
Genomic sequences will be mined with specialized software to develop novel molecular identification and genotyping methods for the insect pathogens Beauveria and Metarhizium. Marker data obtained from strains collected by the scientists, collaborators or acquired from culture collections will form the basis for systematic and taxonomic revisions and the creation of on-line molecular and morphological identification resources. A molecular epidemiological analysis of Beauveria pathogens of coffee berry borer, an important insect pest of coffee, will be conducted in coffee farms in Puerto Rico. Molecular methods for species identification will be critically tested. Population genetic analyses combining microsatellite, mating type and SNP data will be used to analyze the genetic structure of epizootics against the environmental diversity of indigenous Beauveria. In addition, the efficacy and persistence and geographic spread of GHA, the Beauveria mycoinsecticide Mycotrol® strain that has been released for control of coffee berry borer, will be assessed. For the project on pathogenic and endophytic strains of Colletotrichum and Pestalotiopsis, strains will be obtained from a network of cooperators in the Neotropics. Initial diversity screens of Colletotrichum strains will be conducted using markers APN2 and APN2/MATigs; diversity screens for Pestalotiopsis will utilize variable intron regions of elongation factor-1 alpha (EF-1a) and beta tubulin (B-tub). Strains selected from the diversity screens will be used for multi-locus phylogenetic analyses (B-tub, EF-1a, RPB1, RPB2 and other loci) to delineate species boundaries, infer species relationships, and determine whether endophytic and pathogenic ecologies are phylogenetically associated.